Structure and functional mechanisms of molecular chaperones and protein kinases

分子伴侣和蛋白激酶的结构和功能机制

• 批准号: 10552406
• 负责人: CHARALAMPOS KALODIMOS
• 金额: $ 76.43万
• 依托单位: ST. JUDE CHILDREN'S RESEARCH HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-05-01 至 2028-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10552406
• 关键词: Address; Architecture; BRAF gene; Biochemical; Cell physiology; Cells; Cessation of life; Client; Complex; Cytoplasm; Family; Goals; Human; Human Genome; Hyperactivity; Malignant Neoplasms; Metabolism; Molecular Chaperones; Molecular Conformation; Molecular Structure; Mutate; NMR Spectroscopy; Phosphotransferases; Proliferating; Protein Dynamics; Protein Family; Protein Kinase; Protein Tyrosine Kinase; Proteins; Proteome; Reporting; Resolution; Role; Signal Pathway; Structure; Therapeutic Intervention; Work; biophysical techniques; cell growth; member; migration; non-Native; novel; protein aggregation; prototype; raf Kinases; targeted treatment; tumor

PROJECT SUMMARY Our lab works on two main directions: first, the determination of the structural and dynamic basis for the function and assembly of large protein machineries; and second, the determination of the role of internal protein dynamics in regulating protein activity and allosteric interactions. We propose to use NMR spectroscopy, together with other biochemical and biophysical techniques, to determine at the atomic resolution the mechanisms underpinning the function of two important protein families: molecular chaperones and protein kinases. Molecular chaperones are central to maintaining a functional proteome in the cell by rescuing non- native proteins from aggregation and misfolding and assisting with their folding. Our lab reported the first ever high resolution structures of molecular chaperones in complex with unfolded proteins. We will determine the structures of important chaperones such as the Hsp40, Hsp70 and Hsp90 in complex with client proteins. We wish to address how different chaperones engage non-native proteins and how distinct chaperone architectures may alter activity. Src and BRAF hold a prominent place among the over 500 protein kinases encoded by the human genome. Src is the archetypical kinase of the ~30 cytoplasmic tyrosine kinases, half of which (Src, Frk, Abl, Tec and Csk families) share the same core domain architecture consisting of an SH3, SH2 and kinase domain. As such, Src has served as the prototype for understanding how allosteric interactions regulate the activity of kinases. BRAF is a member of RAF kinases and is a part of the ERK signaling pathway, one of the most important and best studied signaling pathways in human cancers. The BRAF gene is mutated in approximately 8% of human tumors with substitutions in the A-loop, the P-loop and the DFG motif reported to give rise to a hyperactive kinase. We will use NMR spectroscopy to characterize transiently populated conformational states, with the goal of uncovering novel regulatory mechanisms in the human kinome.

项目摘要 我们的实验室在两个主要方向上工作：第一，确定结构和动态基础 大型蛋白质机器的功能和组装；其次，确定角色 内部蛋白质动力学控制蛋白质活性和变构相互作用。我们建议使用 NMR光谱以及其他生化和生物物理技术，以确定 原子分辨率是两个重要蛋白质家族功能的机制： 分子伴侣和蛋白激酶。 分子伴侣通过挽救非 - 天然蛋白质来自聚集，错误折叠和协助折叠。我们的实验室报告了 与未折叠的蛋白质复合物中分子伴侣的第一个高分辨率结构。我们 将确定重要伴侣的结构，例如HSP40，HSP70和HSP90 与客户蛋白的复杂性。我们希望解决不同的伴侣如何参与非本地 蛋白质以及不同的伴侣结构如何改变活性。 SRC和BRAF在人类编码的500多个蛋白激酶中占有重要地位 基因组。 SRC是〜30个细胞质酪氨酸激酶的原型激酶，其中一半（src， FRK，ABL，TEC和CSK家庭）共享相同的核心域架构，由SH3，SH2组成 和激酶域。因此，SRC是理解变构的原型 相互作用调节激酶的活性。 Braf是RAF激酶的成员，是 ERK信号通路，人类中最重要，最佳的研究信号通路之一 癌症。 BRAF基因在大约8％的人类肿瘤中突变，并取代 据报道，A环，P环和DFG基序会引起多动激酶。我们将使用NMR 光谱法表征瞬时人口稠密的会议状态，目的是发现 人类神组中的新型调节机制。